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BRAF V600E Mutation testing by RT-PCR
|Performed by||Molecular Diagnostics|
|In House Availability||Run 1x per week, DNA extraction started on Monday, day shift only|
|Method||The BRAF Mutation Testing by Real-time PCR assay utilizes real-time PCR amplification of a portion of exon 15 of the BRAF gene using specific PCR primers. DNA is extracted from tumor tissue and amplified in the real-time PCR reaction. The amplified product is detected with a specific pair of fluorescence resonance energy transfer (FRET) probes which hybridize to the site of potential mutation at nucleotide 1799, corresponding to amino acid 600. After amplification, fluorescence from the probes is measured as the PCR products are heated to cause dissociation of the probes from the PCR products. The temperature at which dissociation occurs is dependent upon the sequence at the site of hybridization.|
|Collection Instructions||Blocks selected for BRAF mutation testing must contain tumor tissue.
Areas of tumor must contain at least 50% tumor cells. The lab can remove adjacent non-tumor tissue, so the entire slide does not need 50% tumor cells, just the area with tumor.
Label slides with pathology case number and block identification.
For UCSF Samples (from remote sites) Click here for sample collection instructions
For NON-UCSF Samples Click here for Requisition form & Account set-up instructions. Note we only do institutional billing.
|Sample type||Formalin-fixed, paraffin-embedded 10 micron tissue sections on five (5) unstained, uncharged glass slides. One adjacent H&E stained slide should be included.|
|UCSF Rejection Criteria||All required slides not included. Insufficient tumor tissue present on slide as determined by pathologist. Slides not labeled or not accompanied by completed requisition form (if not ordered directly through Department of Pathology).|
|Normal range||No mutation detected|
|Stability||Formalin-fixed, paraffin-embedded tissues are stable indefinitely at room temperature|
|Turn around times||10-14 days|
|Reflex?||The test may be performed reflexively on colorectal carcinomas which show both microsatellite instability by PCR, and loss of MLH1 protein by immunohistochemistry.|
|Additional information||This test detects the V600E mutation in the BRAF gene from formalin-fixed paraffin-embedded tumor tissue. BRAF V600E mutation has been shown to be present in some thyroid and colorectal carcinomas, as well as in some melanomas.
In the thyroid, presence of the mutation is associated with more aggressive clinical behavior. BRAF mutation in Lynch syndrome-associated colorectal cancers is extremely rare, and the presence of BRAF mutation is a strong indication of a sporadic (i.e., non-Lynch) colorectal cancer.
Results of this test should be correlated with the patient's other clinical and laboratory information. This assay is not intended to test for BRAF mutations other than the valine to glutamate substitution at amino acid position 600.
The test was validated by UCSF Clinical Laboratories to confirm performance characteristics, in compliance with current guidelines for clinical implementation
|LDT or Mod FDA?||Yes|
|Last Updated||3/13/2013 9:05:05 AM|
If you have additional questions regarding this test, please call: 415-353-1667